This invention relates to immunogenetics and to peptide chemistry. More particularly, it relates to peptides useful in various ways, including immunogens and as ligands for HLA-Cw*16 molecules. More particularly, it relates to so-called xe2x80x9ctumor rejection antigensxe2x80x9d, derived from the tumor rejection antigen precursor encoded by gene MAGE-6, and presented by molecules of MHC class HLA-Cw*16.
The study of the recognition or lack of recognition of cancer cells by a host organism has proceeded in many different directions. Understanding of the field presumes some understanding of both basic immunology and oncology.
Early research on mouse tumors revealed that these displayed molecules which led to rejection of tumor cells when transplanted into syngeneic animals. These molecules are xe2x80x9crecognizedxe2x80x9d by T-cells in the recipient animal, and provoke a cytolytic T-cell response with lysis of the transplanted cells. This evidence was first obtained with tumors induced in vitro by chemical carcinogens, such as methylcholanthrene. The antigens expressed by the tumors and which elicited the T-cell response were found to be different for each tumor. See Prehn, et al., J. Natl. Canc. Inst. 18: 769-778 (1957); Klein et al., Cancer Res. 20: 1561-1572 (1960); Gross, Cancer Res. 3: 326-333 (1943), Basombrio, Cancer Res. 30: 2458-2462 (1970) for general teachings on inducing tumors with chemical carcinogens and differences in cell surface antigens. This class of antigens has come to be known as xe2x80x9ctumor specific transplantation antigensxe2x80x9d or xe2x80x9cTSTAsxe2x80x9d. Following the observation of the presentation of such antigens when induced by chemical carcinogens, similar results were obtained when tumors were induced in vitro via ultraviolet radiation. See Kripke, J. Natl. Canc. Inst. 53: 333-1336 (1974).
While T-cell mediated immune responses were observed for the types of tumor described supra, spontaneous tumors were thought to be generally non-immunogenic. These were therefore believed not to present antigens which provoked a response to the tumor in the tumor carrying subject. See Hewitt, et al., Brit. J. Cancer 33: 241-259 (1976).
The family of tumxe2x88x92 antigen presenting cell lines are immunogenic variants obtained by mutagenesis of mouse tumor cells or cell lines, as described by Boon et al., J. Exp. Med. 152: 1184-1193 (1980), the disclosure of which is incorporated by reference. To elaborate, tumxe2x88x92 antigens are obtained by mutating tumor cells which do not generate an immune response in syngeneic mice and will form tumors (i.e., xe2x80x9ctum+xe2x80x9d cells). When these tum+ cells are mutagenized, they are rejected by syngeneic mice, and fail to form tumors (thus xe2x80x9ctumxe2x88x92xe2x80x9d). See Boon et al., Proc. Natl. Acad. Sci. USA 74: 272 (1977), the disclosure of which is incorporated by reference. Many tumor types have been shown to exhibit this phenomenon. See, e.g., Frost et al., Cancer Res. 43: 125 (1983).
It appears that tumxe2x88x92 variants fail to form progressive tumors because they initiate an immune rejection process. The evidence in favor of this hypothesis includes the ability of xe2x80x9ctumxe2x88x92xe2x80x9d variants of tumors, i.e., those which do not normally form tumors, to do so in mice with immune systems suppressed by sublethal irradiation, Van Pel et al., Proc. Natl. Acad. Sci. USA 76: 5282-5285 (1979); and the observation that intraperitoneally injected tumxe2x88x92 cells of mastocytoma P815 multiply exponentially for 12-15 days, and then are eliminated in only a few days in the midst of an influx of lymphocytes and macrophages (Uyttenhove et al., J. Exp. Med. 152: 1175-1183 (1980)). Further evidence includes the observation that mice acquire an immune memory which permits them to resist subsequent challenge to the same tumxe2x88x92 variant, even when immunosuppressive amounts of radiation are administered with the following challenge of cells (Boon et al., Proc. Natl, Acad. Sci. USA 74: 272-275 (1977); Van Pel et al., supra; Uyttenhove et al., supra). Later research found that when spontaneous tumors were subjected to mutagenesis, immunogenic variants were produced which did generate a response. Indeed, these variants were able to elicit an immune protective response against the original tumor. See Van Pel et al., J. Exp. Med. 157: 1992-2001 (1983). Thus, it has been shown that it is possible to elicit presentation of a so-called xe2x80x9ctumor rejection antigenxe2x80x9d in a tumor which is a target for a syngeneic rejection response. Similar results have been obtained when foreign genes have been transfected into spontaneous tumors. See Fearon et al., Cancer Res. 48: 2975-1980 (1988) in this regard.
A class of antigens has been recognized which are presented on the surface of tumor cells and are recognized by cytolytic T cells, leading to lysis. This class of antigens will be referred to as xe2x80x9ctumor rejection antigensxe2x80x9d or xe2x80x9cTRAsxe2x80x9d hereafter. TRAs may or may not elicit antibody responses. The extent to which these antigens have been studied, has been via cytolytic T cell characterization studies, in vitro i.e., the study of the identification of the antigen by a particular cytolytic T cell (xe2x80x9cCTLxe2x80x9d hereafter) subset. The subset proliferates upon recognition of the presented tumor rejection antigen, and the cells presenting the tumor rejection antigens are lysed. Characterization studies have identified CTL clones which specifically lyse cells expressing the tumor rejection antigens. Examples of this work may be found in Levy et al., Adv. Cancer Res. 24: 1-59 (1977); Boon et al., J. Exp. Med. 152: 1184-1193 (1980); Brunner et al., J. Immunol. 124: 1627-1634 (1980); Maryanski et al., Eur. J. Immunol. 124: 1627-1634 (1980); Maryanski et al., Eur. J. Immunol. 12: 406-412 (1982); Palladino et al., Canc. Res. 47: 5074-5079 (1987). This type of analysis is required for other types of antigens recognized by CTLs, including minor histocompatibility antigens, the male specific H-Y antigens, and the class of antigens referred to as xe2x80x9ctumxe2x88x92xe2x80x9d antigens, and discussed herein.
A tumor exemplary of the subject matter described supra is known as P815. See DePlaen et al., Proc. Natl. Acad. Sci. USA 85: 2274-2278 (1988); Szikora et al., EMBO J 9: 1041-1050 (1990), and Sibille et al., J. Exp. Med. 172: 35-45 (1990), the disclosures of which are incorporated by reference. The P815 tumor is a mastocytoma, induced in a DBA/2 mouse with methylcholanthrene and cultured as both an in vitro tumor and a cell line. The P815 line has generated many tumxe2x88x92 variants following mutagenesis, including variants referred to as P91A (DePlaen, supra), 35B (Szikora, supra), and P198 (Sibille, supra). In contrast to tumor rejection antigensxe2x80x94and this is a key distinctionxe2x80x94the tumxe2x88x92 antigens are only present after the tumor cells are mutagenized. Tumor rejection antigens are present on cells of a given tumor without mutagenesis. Hence, with reference to the literature, a cell line can be tum+, such as the line referred to as xe2x80x9cP1xe2x80x9d, and can be provoked to produce tumxe2x88x92variants. Since the tumxe2x88x92 phenotype differs from that of the parent cell line, one expects a difference in the DNA of tumxe2x88x92 cell lines as compared to their tum+ parental lines, and this difference can be exploited to locate the gene of interest in tumxe2x88x92 cells. As a result, it was found that genes of tumxe2x88x92 variants such as P91A, 35B and P198 differ from their normal alleles by point mutations in the coding regions of the gene. See Szikora and Sibille, supra, and Lurquin et al., Cell 58: 293-303 (1989). This has proved not to be the case with the TRAs of this invention. These papers also demonstrated that peptides derived from the tumxe2x88x92 antigen are presented by the Ld molecule for recognition by CTLs. P91A is presented by Ld, P35 by Dd and P198 by Kd.
PCT application PCT/US92/04354, filed on May 22, 1992 assigned to the same assignee as the subject application, teaches a family of human tumor rejection antigen precursor coding genes, referred to as the MAGE family. Several of these genes are also discussed in van der Bruggen et al., Science 254: 1643 (1991). It is now clear that the various genes of the MAGE family are expressed in tumor cells, and can serve as markers for the diagnosis of such tumors, as well as for other purposes discussed therein. See also Traversari et al., Immunogenetics 35: 145 (1992); van der Bruggen et al., Science 254: 1643 (1991) and De Plaen, et al., Immunogenetics 40: 360 (1994). The mechanism by which a protein is processed and presented on a cell surface has now been fairly well documented. A cursory review of the development of the field may be found in Barinaga, xe2x80x9cGetting Some xe2x80x98Backbonexe2x80x99: How MHC Binds Peptidesxe2x80x9d, Science 257: 880 (1992) also, see Fremont et al., Science 257: 919 (1992); Matsumura et al., Science 257: 927 (1992); Latron et al., Science 257: 964 (1992). These papers generally point to a requirement that the peptide which binds to an MHC/HLA molecule be nine amino acids long (a xe2x80x9cnonapeptidexe2x80x9d), and to the importance of the first and ninth residues of the nonapeptide.
Studies on the MAGE family of genes have now revealed that a particular nonapeptide is in fact presented on the surface of some tumor cells, and that the presentation of the nonapeptide requires that the presenting molecule be HLA-A1. Complexes of the MAGE-1 tumor rejection antigen (the xe2x80x9cTRAxe2x80x9d or nonapeptidexe2x80x9d) leads to lysis of the cell presenting it by cytolytic T cells (xe2x80x9cCTLsxe2x80x9d).
Attention is drawn, e.g., to applications Ser. No. 08/217,188 now U.S. Pat. No. 5,554,724 to Melief et al, Ser. No. 08/217,187 now U.S. Pat. No. 5,554,506 to Traversari et al., and Ser. No. 08/217,186 now U.S. Pat. No. 5,535,461 to Townsend et al., all of which present work on other, MAGE-derived peptides.
Research presented in, e.g., U.S. patent application Ser. No. 07/938,334 filed Aug. 31, 1992 now U.S. Pat. No. 5,404,940, and in U.S. patent application Ser. No. 073,103, filed Jun. 7, 1993 now U.S. Pat. No. 5,462,871, found that when comparing homologous regions of various MAGE genes to the region of the MAGE-1 gene coding for the relevant nonapeptide, there is a great deal of homology. Indeed, these observations lead to one of the aspects of the invention disclosed and claimed therein, which is a family of nonapeptides all of which have the same N-terminal and C-terminal amino acids. These nonapeptides were described as being useful for various purposes which includes their use as immunogens, either alone or coupled to carrier peptides. Nonapeptides are of sufficient size to constitute an antigenic epitope, and the antibodies generated thereto were described as being useful for identifying the nonapeptide, either as it exists alone, or as part of a larger polypeptide.
The human major histocompatibility complex (MHC) system is an involved one. One feature of the system is the human leukocyte antigen, or xe2x80x9cHLAxe2x80x9d. Human cells can be xe2x80x9ctypedxe2x80x9d, based upon their HLA profile. Not all cells present all types of HLA molecules. The diversity of this system may be seen by reference to, e.g., Zemmour et al., Immunogenetics 37: 239-250 (1993). This reference shows that, as of 1992, there were literally dozens of different HLA alleles which were known to the art. Cianetti et al., Immunogenetics 29: 80-91 (1989), the disclosure of which is incorporated by reference, discloses, inter alia, an MHC allele referred to therein as HLA-C-clone 10. This allele was later renamed, as per Bodmer et al., Tissue Antigen 44: 1-18 (1994), incorporated by reference. The allele is now known as HLA-Cw*1601. A related allele, i.e., HLA-Cw*1602 is also known. See Bodmer et al, supra; Vilches et al., Human Immunol. 41: 167-170 (1994), also incorporated by reference. These two alleles, when discussed collectively, will be referred to as xe2x80x9cHLA-Cw*16xe2x80x9d. Van der Bruggen et al, Eur. J. Immunol. 24: 2134-2140 (1994), incorporated by reference, teach that cytolytic T cells (xe2x80x9cCTLsxe2x80x9d) recognize a complex of a peptide and an HLA-Cw*1601 molecule. These data are also disclosed in copending U.S. patent application Ser. No. 08/79,110, filed Jun. 17, 1993, now U.S. Pat. No. 5,571,711 and incorporated by reference, and in U.S. patent application Ser. No. 08/196,630, filed on Feb. 15, 1994 U.S. Pat. No. 5,683,886, and incorporated by reference. Both applications are assigned to the assignee of the subject application. These two applications disclose that the tumor rejection antigen precursor referred to as xe2x80x9cBAGExe2x80x9d is processed to peptides presented by the HLA-Cw*1601 molecule. Specifically, the applications disclose a preferred nonapeptide:
Ala Ala Arg Ala Val Phe Leu Ala Leuxe2x80x83xe2x80x83(SEQ ID NO: 1),
which complexes with HLA-Cw*1601, thereby stimulating the cell line CTL 82/82.
U.S. Pat. No. 5,342,774, incorporated by reference, discloses a family of related nucleic acid molecules which encode tumor rejection antigen precursors referred to as the MAGE tumor rejection antigen precursor. These xe2x80x9cTRAPsxe2x80x9d are numbered as MAGE-1, MAGE-2, etc. Generally, they are expressed predominantly on tumor cells, normal testes cells being the major exception, and they are processed to peptides presented by various HLA molecules, such as HLA-A1, HLA-A2, and so forth. See, e.g., U.S. Pat. No. 5,405,940, incorporated by reference.
Not all tumor cells express all MAGE TRAPs. Hence, while it is desirable for the field to have information available on specific complexes of peptide and HLA molecules which do stimulate T cells, this is not always easy to secure. One requires a cell which expresses both the required MHC molecule and the required TRAP molecule, and the ability to process the large TRAP molecule to the smaller, tumor rejection antigen or xe2x80x9cTRAxe2x80x9d. Even when a particular peptide binds with an MHC molecule to form a complex, this does not per se guarantee that the complex will stimulate CTL proliferation. In other words while peptide MHC binding is sufficient to identify MHC phenotype on a cell, this binding is necessary but not sufficient to provoke CTL proliferation.
The inventors have found, however, that the TRAP encoded by the MAGE-6 gene is processed to TRAs which bind to MHC Molecules of type HLA-Cw*16, and also provoke proliferation of CTLs thereby. Hence, the invention is directed, inter alia, to these peptides and their various uses, as will be explained in the disclosure which follows.